The fluorescence characteristic of small molecular organic compound in nature has potential application value for plant species identification. At present, the identification of two species about Dalbergia cochinchinensis Pierre and Dalbergia oliveri Prain based on the anatomical characteristics of wood has its limitation. So, we explored a new method for identification of Dalbergia cochinchinensis Pierre and Dalbergia oliveri Prain by fluorescence spectroscopy in the paper. The fluorescent substances were completely extracted in five different wood samples of Dalbergia cochinchinensis Pierre and Dalbergia oliveri Prain. Fluorescence photos, two-dimensional and three-dimensional fluorescence spectrum were used to confirm the species and quantity of fluorescent substances in wood samples of the two species. The results of fluorescence photograph shown that the extract solutions of Dalbergia cochinchinensis Pierre and Dalbergia oliveri Prain both present obvious fluorescence characteristics and the fluorescence substances are stable in two years of storage. Two-dimensional and three-dimensional fluorescence spectrum exhibit the fluorescence characteristics of the two species in more detail. The results indicate that there is a fluorescent substance in the extract solution of Dalbergia cochinchinensis Pierre, and the maximum excitation and emission wavelength of the fluorescence substance are 297 and 327 nm, respectively. However, the fluorescence spectrum of extract solution from Dalbergia oliveri Prain present two maximal excitation and emission wavelengths respectively, the maximum excitation is 317 and 333 nm, and the maximum emission is 342 and 358 nm, which indicates that there are two kinds of fluorescent substances or one kind of fluorescence substance containing two groups of fluorescence. The respective quantity and characteristics of fluorescent substances of the two species are obviously accordant, and there are obvious differences between the two species, which can be used to confirm their identities. Moreover, the extracting methods of fluorescent substances from the wood sample of the two species are built in this paper. Acetonitrile is an optimal reagent for the extraction of fluorescent substance in the wood sample of Dalbergia cochinchinensis Pierre, and the mixed reagent prepared according to the volume ratio of ethyl acetate and acetonitrile equals 10∶1 is more suitable for that of fluorescent substance in Dalbergia oliveri Prain. The extraction of fluorescent substance is beginning to level off within 12 hours for the wood samples of Dalbergia cochinchinensis Pierre and Dalbergia oliveri Prain. The results indicate that the fluorescence spectroscopy technique can be used to identify Dalbergia cochinchinensis Pierre and Dalbergia oliveri Prain. The method will be of very important value to the identification of rosewood species.